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Toxicological information

Genetic toxicity: in vitro

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Administrative data

Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Study period:
1991
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Well performed and reported GLP study; however using only four Salmonella strains TA 98, 100, 1535 and 1537 and not the E coli strain (not required in 1992).

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1992
Report Date:
1992

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
other: solution
Details on test material:
Name: Trilon B-CU 1550
Batch no.: Labor Journal No. 91 233
Test substance no.: 91/401
Purity: 47.2% (52.1% water)
Appearance: blue liquid

Method

Target gene:
Point mutation
Species / strain
Species / strain / cell type:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Additional strain / cell type characteristics:
not applicable
Metabolic activation:
with and without
Metabolic activation system:
Aroclor 1254 induced rat liver S9 mix
Test concentrations with justification for top dose:
First (standard plate test) and second (preincubation) test: 0, 100, 500, 2500, 5000 and 10000 µg/plate
Vehicle / solvent:
water
Controls
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
True negative controls:
no
Positive controls:
yes
Positive control substance:
9-aminoacridine
other: 2-aminoanthracene, N-methyl-N-nitro-N-nitrososguanidine, nitro-o-phenylendiamine
Remarks:
2-AA was used with S9-mix, the other without S9-mix
Details on test system and experimental conditions:
METHOD OF APPLICATION: standard plate test (Ames) and preincubation test (Yagahi et al. and Matsushima et al.)

DURATION
- Preincubation period: 20 min
- Exposure duration: 48 h

NUMBER OF REPLICATIONS: 3 test plates per dose

NUMBER OF CELLS EVALUATED: colonies are counted

DETERMINATION OF CYTOTOXICITY
- Method: his- background growth

Evaluation criteria:
Positive result in case of:
- doubling of the spontaneous mutation rate (control)
- dose-response relationship
- reproducibility of the results
Statistics:
No info

Results and discussion

Test results
Species / strain:
S. typhimurium TA 1535, TA 1537, TA 98 and TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity nor precipitates, but tested up to recommended limit concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not examined
Positive controls validity:
valid
Additional information on results:
See for test results the Tables below
Remarks on result:
other: strain/cell type:
Remarks:
Migrated from field 'Test system'.

Any other information on results incl. tables

Results of Strain TA 1535 (standard plate)

 

Dose (ug/plate)

Number of revertants per plate

-S9-mix

+S9-mix

Mean

SD

Mean

SD

0

19

3

22

3

100

17

5

22

2

500

18

3

22

2

2500

22

4

24

3

5000

17

2

22

1

10000

17

3

13

0

2-AA

-

-

145

7

MNNG

1017

6

-

-

 

Results of Strain TA 100 (standard plate)

 

Dose (ug/plate)

Number of revertants per plate

-S9-mix

+S9-mix

Mean

SD

Mean

SD

0

124

11

144

6

100

124

21

149

4

500

134

19

214

15

2500

124

21

152

16

5000

131

6

133

10

10000

126

17

146

4

2-AA

-

-

1173

170

MNNG

1037

96

-

-

 

Results of Strain TA 1537 (standard plate)

 

Dose (ug/plate)

Number of revertants per plate

-S9-mix

+S9-mix

Mean

SD

Mean

SD

0

8

2

10

3

100

8

3

12

2

500

8

2

14

2

2500

8

3

10

1

5000

9

1

9

4

10000

7

4

9

1

2-AA

-

-

116

6

AAC

563

46

-

-

 

Results of Strain TA 98 (standard plate)

 

Dose (ug/plate)

Number of revertants per plate

-S9-mix

+S9-mix

Mean

SD

Mean

SD

0

18

3

32

4

100

16

6

33

7

500

22

4

32

2

2500

22

1

26

6

5000

23

2

18

3

10000

24

1

23

7

2-AA

-

-

738

58

NPD

688

32

-

-

 

Results of Strain TA 1535 (preincubation)

 

Dose (ug/plate)

Number of revertants per plate

-S9-mix

+S9-mix

Mean

SD

Mean

SD

0

16

4

13

3

100

14

2

11

1

500

15

3

9

1

2500

12

3

11

3

5000

14

5

11

3

10000

11

5

11

2

2-AA

-

-

82

3

MNNG

571

30

-

-

 

Results of Strain TA 100 (preincubation)

 

Dose (ug/plate)

Number of revertants per plate

-S9-mix

+S9-mix

Mean

SD

Mean

SD

0

100

14

107

5

100

95

12

106

3

500

98

15

124

5

2500

123

5

107

8

5000

107

14

126

3

10000

109

12

128

19

2-AA

-

-

561

9

MNNG

578

66

-

-

 

Results of Strain TA 1537 (preincubation)

 

Dose (ug/plate)

Number of revertants per plate

-S9-mix

+S9-mix

Mean

SD

Mean

SD

0

8

1

12

2

100

8

3

9

1

500

7

3

9

2

2500

6

2

7

2

5000

7

0

6

2

10000

5

3

8

0

2-AA

-

-

78

9

AAC

412

14

-

-

 

Results of Strain TA 98 (preincubation)

 

Dose (ug/plate)

Number of revertants per plate

-S9-mix

+S9-mix

Mean

SD

Mean

SD

0

28

5

27

7

100

22

3

29

2

500

22

2

26

9

2500

17

1

25

2

5000

26

3

24

1

10000

25

2

23

2

2-AA

-

-

556

39

NPD

744

38

-

-

 

 

Applicant's summary and conclusion

Conclusions:
Interpretation of results (migrated information):
negative

The test substance EDTA-CuNa2 is not mutagenic in the Ames test under the experiemnatl conditions used.
Executive summary:

The test substance EDTA-CuNa2 (Trilon B-CU 1550) was tested for its mutagenic potential based on the ability to induce back mutations in selected loci in several strains of Salmonelle thyphimurium (TA 1535, TA 100, TA 1537 and TA 98) in the Ames test. The dose range was between 100 and 10000 ug/plate. The standard plate test and the preincubation test were used both with and without metabolic activation (Aroclor induced rat liver S9 -mix). No precipitation of the test substance was found. No bacteriotoxic effect was observed. An increase in the number of His+ revertants was not observed both in the standard plate test and in the pre-incubation test either with or without S9 -mix. Based on the results of this study, the test substance EDTA-CuNa2 is not mutagenic in the Ames test under the experiemnatl conditions used.